FIGURE 1.
Analysis of AcnR-DNA interaction by EMSAs. A, determination of the apparent KD for the binding of AcnR to its operator in the acn promoter. A 2 nm concentration of a Cy3-labeled DNA fragment covering the promoter region of acn was incubated with increasing concentrations of AcnR and analyzed as described under “Experimental Procedures.” An apparent KD of 7.7 ± 0.9 nm was determined for dimeric AcnR. B, competition experiment with specific and nonspecific DNA (unlabeled), 2 nm Cy3-labeled DNA of the acn promoter, and 25 nm AcnR dimer. Specific DNA was unlabeled acn promoter DNA (10 nm, 40 nm, 100 nm), and nonspecific DNA was a 468-bp fragment covering the promoter region of cg1848 (10, 40, and 100 nm).